Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism

Yong Gang Chang; Christopher J. Lupton; Charles Bayly-Jones; Alastair C. Keen; Laura D’Andrea; Christina M. Lucato; Joel R. Steele; Hari Venugopal; Ralf B. Schittenhelm; James C. Whisstock; Michelle L. Halls; Andrew M. Ellisdon

doi:10.1038/s41594-022-00804-9

Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism

Yong Gang Chang, Christopher J. Lupton, Charles Bayly-Jones, Alastair C. Keen, Laura D’Andrea, Christina M. Lucato, Joel R. Steele, Hari Venugopal, Ralf B. Schittenhelm, James C. Whisstock, Michelle L. Halls, Andrew M. Ellisdon

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5 Citations (Scopus)

Abstract

P-Rex (PI(3,4,5)P₃-dependent Rac exchanger) guanine nucleotide exchange factors potently activate Rho GTPases. P-Rex guanine nucleotide exchange factors are autoinhibited, synergistically activated by Gβγ and PI(3,4,5)P₃ binding and dysregulated in cancer. Here, we use X-ray crystallography, cryogenic electron microscopy and crosslinking mass spectrometry to determine the structural basis of human P-Rex1 autoinhibition. P-Rex1 has a bipartite structure of N- and C-terminal modules connected by a C-terminal four-helix bundle that binds the N-terminal Pleckstrin homology (PH) domain. In the N-terminal module, the Dbl homology (DH) domain catalytic surface is occluded by the compact arrangement of the DH-PH-DEP1 domains. Structural analysis reveals a remarkable conformational transition to release autoinhibition, requiring a 126° opening of the DH domain hinge helix. The off-axis position of Gβγ and PI(3,4,5)P₃ binding sites further suggests a counter-rotation of the P-Rex1 halves by 90° facilitates PH domain uncoupling from the four-helix bundle, releasing the autoinhibited DH domain to drive Rho GTPase signaling.

Original language	English
Pages (from-to)	767–773
Number of pages	25
Journal	Nature Structural & Molecular Biology
Volume	29
Issue number	8
DOIs	https://doi.org/10.1038/s41594-022-00804-9
Publication status	Published - Aug 2022

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10.1038/s41594-022-00804-9Licence: CC BY

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Chang, Y. G., Lupton, C. J., Bayly-Jones, C., Keen, A. C., D’Andrea, L., Lucato, C. M., Steele, J. R., Venugopal, H., Schittenhelm, R. B., Whisstock, J. C., Halls, M. L., & Ellisdon, A. M. (2022). Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism. Nature Structural & Molecular Biology, 29(8), 767–773. https://doi.org/10.1038/s41594-022-00804-9

@article{4d8a1800f46e470d9346d7d2af4d56ad,

title = "Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism",

abstract = "P-Rex (PI(3,4,5)P3-dependent Rac exchanger) guanine nucleotide exchange factors potently activate Rho GTPases. P-Rex guanine nucleotide exchange factors are autoinhibited, synergistically activated by Gβγ and PI(3,4,5)P3 binding and dysregulated in cancer. Here, we use X-ray crystallography, cryogenic electron microscopy and crosslinking mass spectrometry to determine the structural basis of human P-Rex1 autoinhibition. P-Rex1 has a bipartite structure of N- and C-terminal modules connected by a C-terminal four-helix bundle that binds the N-terminal Pleckstrin homology (PH) domain. In the N-terminal module, the Dbl homology (DH) domain catalytic surface is occluded by the compact arrangement of the DH-PH-DEP1 domains. Structural analysis reveals a remarkable conformational transition to release autoinhibition, requiring a 126° opening of the DH domain hinge helix. The off-axis position of Gβγ and PI(3,4,5)P3 binding sites further suggests a counter-rotation of the P-Rex1 halves by 90° facilitates PH domain uncoupling from the four-helix bundle, releasing the autoinhibited DH domain to drive Rho GTPase signaling.",

author = "Chang, {Yong Gang} and Lupton, {Christopher J.} and Charles Bayly-Jones and Keen, {Alastair C.} and Laura D{\textquoteright}Andrea and Lucato, {Christina M.} and Steele, {Joel R.} and Hari Venugopal and Schittenhelm, {Ralf B.} and Whisstock, {James C.} and Halls, {Michelle L.} and Ellisdon, {Andrew M.}",

note = "Funding Information: C.B.-J. acknowledges the support of the Australian Government by way of an RTP stipend. M.L.H. is a Viertel Senior Medical Research Fellow supported by The Cross Family and The Frank Alexander Charitable Trusts. J.C.W. is an Australian Research Council Laureate Fellow and honorary National Health and Medical Research Council of Australia Senior Principal Research Fellow. A.M.E. is Victorian Cancer Agency Mid-Career Fellow supported by the Victorian Department of Health and Human Services acting through the VCA. This research was supported by NHMRC Project Grants to A.M.E. (APP1146578, APP1128120), and equipment funded by an Australian Research Council Grant LE170100016. This research was undertaken in part using the MX2 beamline at the Australian Synchrotron—part of ANSTO—and made use of the Australian Cancer Research Foundation (ACRF) detector. The authors acknowledge the use of instruments and assistance at the Monash Ramaciotti Centre for Cryo-Electron Microscopy, a Node of Microscopy Australia. We also acknowledge the office of the Vice-Provost for Research and Research Infrastructure (VPRRI) at Monash University and of Bioplatforms Australia (BPA) as part of the National Collaborative Research Infrastructure Strategy (NCRIS). Publisher Copyright: {\textcopyright} 2022, Crown.",

year = "2022",

month = aug,

doi = "10.1038/s41594-022-00804-9",

language = "English",

volume = "29",

pages = "767–773",

journal = "Nature Structural & Molecular Biology",

issn = "1545-9993",

publisher = "Nature Publishing Group",

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Chang, YG , Lupton, CJ , Bayly-Jones, C , Keen, AC, D’Andrea, L, Lucato, CM, Steele, JR, Venugopal, H, Schittenhelm, RB , Whisstock, JC , Halls, ML & Ellisdon, AM 2022, 'Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism', Nature Structural & Molecular Biology, vol. 29, no. 8, pp. 767–773. https://doi.org/10.1038/s41594-022-00804-9

TY - JOUR

T1 - Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism

AU - Chang, Yong Gang

AU - Lupton, Christopher J.

AU - Bayly-Jones, Charles

AU - Keen, Alastair C.

AU - D’Andrea, Laura

AU - Lucato, Christina M.

AU - Steele, Joel R.

AU - Venugopal, Hari

AU - Schittenhelm, Ralf B.

AU - Whisstock, James C.

AU - Halls, Michelle L.

AU - Ellisdon, Andrew M.

N1 - Funding Information: C.B.-J. acknowledges the support of the Australian Government by way of an RTP stipend. M.L.H. is a Viertel Senior Medical Research Fellow supported by The Cross Family and The Frank Alexander Charitable Trusts. J.C.W. is an Australian Research Council Laureate Fellow and honorary National Health and Medical Research Council of Australia Senior Principal Research Fellow. A.M.E. is Victorian Cancer Agency Mid-Career Fellow supported by the Victorian Department of Health and Human Services acting through the VCA. This research was supported by NHMRC Project Grants to A.M.E. (APP1146578, APP1128120), and equipment funded by an Australian Research Council Grant LE170100016. This research was undertaken in part using the MX2 beamline at the Australian Synchrotron—part of ANSTO—and made use of the Australian Cancer Research Foundation (ACRF) detector. The authors acknowledge the use of instruments and assistance at the Monash Ramaciotti Centre for Cryo-Electron Microscopy, a Node of Microscopy Australia. We also acknowledge the office of the Vice-Provost for Research and Research Infrastructure (VPRRI) at Monash University and of Bioplatforms Australia (BPA) as part of the National Collaborative Research Infrastructure Strategy (NCRIS). Publisher Copyright: © 2022, Crown.

PY - 2022/8

Y1 - 2022/8

N2 - P-Rex (PI(3,4,5)P3-dependent Rac exchanger) guanine nucleotide exchange factors potently activate Rho GTPases. P-Rex guanine nucleotide exchange factors are autoinhibited, synergistically activated by Gβγ and PI(3,4,5)P3 binding and dysregulated in cancer. Here, we use X-ray crystallography, cryogenic electron microscopy and crosslinking mass spectrometry to determine the structural basis of human P-Rex1 autoinhibition. P-Rex1 has a bipartite structure of N- and C-terminal modules connected by a C-terminal four-helix bundle that binds the N-terminal Pleckstrin homology (PH) domain. In the N-terminal module, the Dbl homology (DH) domain catalytic surface is occluded by the compact arrangement of the DH-PH-DEP1 domains. Structural analysis reveals a remarkable conformational transition to release autoinhibition, requiring a 126° opening of the DH domain hinge helix. The off-axis position of Gβγ and PI(3,4,5)P3 binding sites further suggests a counter-rotation of the P-Rex1 halves by 90° facilitates PH domain uncoupling from the four-helix bundle, releasing the autoinhibited DH domain to drive Rho GTPase signaling.

AB - P-Rex (PI(3,4,5)P3-dependent Rac exchanger) guanine nucleotide exchange factors potently activate Rho GTPases. P-Rex guanine nucleotide exchange factors are autoinhibited, synergistically activated by Gβγ and PI(3,4,5)P3 binding and dysregulated in cancer. Here, we use X-ray crystallography, cryogenic electron microscopy and crosslinking mass spectrometry to determine the structural basis of human P-Rex1 autoinhibition. P-Rex1 has a bipartite structure of N- and C-terminal modules connected by a C-terminal four-helix bundle that binds the N-terminal Pleckstrin homology (PH) domain. In the N-terminal module, the Dbl homology (DH) domain catalytic surface is occluded by the compact arrangement of the DH-PH-DEP1 domains. Structural analysis reveals a remarkable conformational transition to release autoinhibition, requiring a 126° opening of the DH domain hinge helix. The off-axis position of Gβγ and PI(3,4,5)P3 binding sites further suggests a counter-rotation of the P-Rex1 halves by 90° facilitates PH domain uncoupling from the four-helix bundle, releasing the autoinhibited DH domain to drive Rho GTPase signaling.

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DO - 10.1038/s41594-022-00804-9

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JO - Nature Structural & Molecular Biology

JF - Nature Structural & Molecular Biology

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Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism

Abstract

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Other files and links

Structural characterisation of the co-inhibitory complex formed by the tumour suppressor PTEN and the metastatic factor PREX2

Structural and functional characterisation of the oncogene P-Rex1

Australian Synchrotron

Centre for Electron Microscopy (MCEM)

Monash Proteomics & Metabolomics Facility

Cite this

Structure of the metastatic factor P-Rex1 reveals a two-layered autoinhibitory mechanism

Abstract

UN SDGs

Access to Document

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Projects

Structural characterisation of the co-inhibitory complex formed by the tumour suppressor PTEN and the metastatic factor PREX2

Structural and functional characterisation of the oncogene P-Rex1

Equipment

Australian Synchrotron

Centre for Electron Microscopy (MCEM)

Monash Proteomics & Metabolomics Facility

Cite this